Peploe grew up in Edinburgh and began his art education after attempting various careers, including working in a solicitors’ office. He enrolled at the Académie Julian in Paris in 1891, and returned there for periods until 1894. He also attended classes at the Royal Scottish Academy between 1892 and 1896. Peploe began his professional career in 1896 when he acquired a studio and began sending work to the annual exhibitions of the Royal Scottish Academy and Royal Glasgow Institute.
Throughout his career, Peploe maintained a studio practice, focussing on the genres of the still life and figure study, as well as painting landscapes en plein air. A trip to Holland developed his interest in the work of seventeenth-century Dutch Old Masters, including Frans Hals and Rembrandt. Their influence, as well as their familiarity with the 19th century French artist Edouard Manet, is apparent in the sophisticated still lifes and figure studies of the late 1890s and early 1900s.
In 1894 Peploe made the first of several painting trips to the island of Barra where he met his future wife Margaret Mackay. Two years later he painted for the first time in North Berwick, a coastal town south of Edinburgh. Using small, transportable panels, these more spontaneous works contrast with the disciplined studio paintings. In around 1900 Peploe became friends with J. D. Fergusson and in 1904 they began to paint together in France each summer.
Peploe made his name with his first solo exhibition, held at The Scottish Gallery in 1903, at which 20 works were sold.